西农萨能奶山羊SERPINA1基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2020.07.001

Abstract

Abstract: The objective of this study was to clone the CDS region of SERPINA1 gene in Xinong Saanen dairy goats (Capra hircus),conduct the bioinformatics analysis using biological software and online prediction tools,and detect the mRNA expression level of SERPINA1 gene in different tissues of Xinong Saanen dairy goats by Real-time quantitative PCR.Specific primers were designed by Primer Premier 5.0 according to the CDS sequence of SERPINA1 gene in Capra hircus (GenBank accession No.:XM_018066209.1),the SERPINA1 gene CDS was amplified by RT-PCR and analyzed by bioinformatics after prokaryotic expression vector was constructed.Collecting tissues such as heart,liver,spleen,lung,mammary gland,kidney,muscle,rumen and small intestine,mRNA was extracted from these tissues and reversed transcription as cDNA,specific primers were designed for Real-time quantitative PCR to detect the expression differences of SERPINA1 gene mRNA in different tissues.The results showed that the CDS region of SERPINA1 gene in Xinong Saanen dairy goats (Capra hircus) was 1 326 bp in length,encoding 441 amino acids.Homology alignment analysis showed that the nucleotide sequences of Xinong Saanen dairy goats with Capra hircus,Ovis aries,Bos taurus and Mus musculus were 100%,98.6%,95.7% and 71.6%,respectively,and the goat was the most closest species with dairy goat,the second was sheep.The SERPINA1 protein had the molecular weight of 48.71 ku,the isoelectric point was 5.71,and was a hydrophilic protein.Amino acid sequence of SERPINA1 had 62 phosphorylation sites,with 3 transmembrane structure.Tissue expression analysis showed that SERPINA1 gene was highly expressed in liver of Xinong Saanen dairy goats (P<0.05),followed by the mammary gland,and was the least expressed in lung.The results provided a theoretical basis for further exploration of the role of SERPINA1 gene in milk protein synthesis and metabolism of dairy goats.

Key words: Xinong Sannen dairy goats; SERPINA1 gene; cloning; bioinformatics analysis; tissue expression

CLC Number:

PAN Tan, GAO Liangjiahui, CHENG Fei, LUO Jun, LI Cong. Cloning,Bioinformatics and Tissue Expression Analysis of SERPINA1 Gene in Xinong Saanen Dairy Goats (Capra hircus)[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(7): 1971-1980.

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Cloning,Bioinformatics and Tissue Expression Analysis of SERPINA1 Gene in Xinong Saanen Dairy Goats (Capra hircus)

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